扑草净胁迫对四角蛤蜊解毒相关基因表达的影响
Prometryn Stress in Expression of Detoxification-related Genes in Mactra veneriformis
-
摘要: 扑草净是我国常用除草剂的主要成分，其药物残留在近岸海域广泛分布。在水温(15±1) ℃条件下，采用半静态水质接触染毒法开展胁迫实验。以β-肌动蛋白(β-Actin)基因为参比，分别研究了不同浓度扑草净胁迫对四角蛤蜊消化腺和鳃组织中超氧化物歧化酶(superoxide dismutase, SOD)、过氧化氢酶(catalase, CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)、谷胱甘肽S-转移酶(glutathione S-transferase, GST)、细胞色素P450第四亚族(cytochrome P450, CYP4)和金属硫蛋白(metallothionein, MT)6种解毒相关基因表达的影响规律。结果显示，6种基因表达调控对扑草净胁迫反应灵敏，在扑草净质量浓度分别为0.2、1.0和10 μg·L-1的养殖海水中，随着胁迫时间延长，四角蛤蜊消化腺呈现SOD、CAT、GSH-Px、GST和MT等5种基因表达均先上调、CYP4基因表达先下调，而后反复波动变化的趋势；鳃呈现SOD、GSH-Px和GST等3种基因表达均先上调，CYP4基因表达先下调，1.0 μg·L-1组的CAT基因表达先下调其他组先上调、MT基因表达先上调其他组先下调，而后反复波动变化的趋势。GST可作为扑草净对四角蛤蜊污染潜在的生物标志物。该结果为评价扑草净残留对海洋生态环境安全风险提供了参考依据。Abstract: Prometryn is a herbicide that is widely used in China and frequently detected in coastal environment. In this study, Mactra veneriformis were exposed to seawaters with prometryn at 0, 0.2, 1.0 μg·L-1, and up to 10.0 μg·L-1 by the semi-static method at (15±1) ℃. Digestive glands and gills in each group of Mactra veneriformis were collected at exposure time 6, 24, 48, 96, 144, 240, 360 and 504 h respectively. The expression of detoxification-related genes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), cytochrome P450 (CYP4) and metallothionein (MT), were analyzed in relative quantification by reverse transcription, quantitative real-time polymerase chain reaction (qPCR) technology and 2-△△ct method with β-Actin gene as the reference gene. The results showed that the regulation of expression of these genes was sensitive to the prometryn and fluctuated over time. The expression of SOD, CAT, GSH-Px, GST, MT mRNA was increased and CYP4 mRNA decreased in the digestive glands of Mactra veneriformis at exposure time 6 h. Subsequently, the expression of these genes was fluctuated in the digestive glands of Mactra veneriformis in all 3 treatment groups. In the gill of Mactra veneriformis, the expression levels of SOD, GSH-Px, GST mRNA increased and CYP4 mRNA decreased in all 3 treated groups at 6 h, when the expression levels of CAT mRNA decreased in the 1.0 μg·L-1 exposure group and increased in other groups. Meanwhile, the expression levels of MT mRNA was increased in the 1.0 μg·L-1 exposure group and decreased in other groups at 6 h, and then fluctuated. GST can be used as a potential mRNA biomarker for the impact of prometryn on Mactra veneriformis. The results provide a promising way for the evaluation of the risk of prometryn residue on marine ecological environment safety.
-
Key words:
- prometryn /
- Mactra veneriformis /
- gene expression /
- detoxification
-
-
United States Environmental Protection Agency (US EPA). Guidance for the reregistration of pesticide products containing prometryn as the active ingredient[R]. Washington DC:US EPA, 1987 European Commission. Commission Regulation (EC) No. 2076/2002[EB/OL]. (2002-11-23)[2021-07-21]. https://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=CELEX:02002R2076-20140715 张树林, 邢克智, 张树伟. 养殖水体水华发生及控制的研究[J]. 水利渔业, 2006, 27(2):67-69 Zhang S L, Xing K Z, Zhang S W. Research on the occurrence and control of aquaculture water[J]. Reservoir Fisheries, 2006, 27(2):67-69(in Chinese)
中华人民共和国农业部. 中华人民共和国农业部公告1435号[EB/OL]. (2010-07-30 )[2021-07-21]. http://www.foodmate.net/law/shipin/164493.html
张骞月, 吴伟. 扑草净在养殖水体中的生态毒理效应及其微生物降解的研究进展[J]. 生物灾害科学, 2014, 37(1):64-69 Zhang Q Y, Wu W. Research advance in eco-toxical effects and microbial degradation of prometryne in aquaculture water[J]. Biological Disaster Science, 2014, 37(1):64-69(in Chinese)
Yang L Q, Li H M, Zhang Y Y, et al. Environmental risk assessment of triazine herbicides in the Bohai Sea and the Yellow Sea and their toxicity to phytoplankton at environmental concentrations[J]. Environment International, 2019, 133(Pt A):105175 Dsikowitzky L, Nguyen T, Konzer L, et al. Occurrence and origin of triazine herbicides in a tropical coastal area in China:A potential ecosystem threat[J]. Estuarine, Coastal and Shelf Science, 2020, 235:106612 徐英江, 刘慧慧, 任传博, 等. 莱州湾海域表层海水中三嗪类除草剂的分布特征[J]. 渔业科学进展, 2014, 35(3):34-39 Xu Y J, Liu H H, Ren C B, et al. Distributions of the triazine herbicides in the surface seawater of Laizhou Bay[J]. Progress in Fishery Sciences, 2014, 35(3):34-39(in Chinese)
Munaron D, Tapie N, Budzinski H, et al. Pharmaceuticals, alkylphenols and pesticides in Mediterranean coastal waters:Results from a pilot survey using passive samplers[J]. Estuarine, Coastal and Shelf Science, 2012, 114:82-92 Mai C. Spatial, seasonal and vertical distributions of currently-used pesticides in the marine boundary layer of the North Sea[J]. Atmospheric Environment, 2013, 75:92-102 李庆鹏, 秦达, 崔文慧, 等. 我国水产品中农药扑草净残留超标的警示分析[J]. 食品安全质量检测学报, 2014, 5(1):108-112 Li Q P, Qin D, Cui W H, et al. Warning analysis on excess prometryne residues of aquatic products in China[J]. Journal of Food Safety & Quality, 2014, 5(1):108-112(in Chinese)
食品资讯中心. 2019年7月输日食品违反日本食品卫生法情况(7月汇总)[EB/OL]. (2019-08 -05)[2021-07-23]. http://news.foodmate.net/2019/08/528758.html
邹婉虹, 刘露, 孙涛, 等. 扑草净在我国养殖水产品中的残留及风险评估[J]. 中国渔业质量与标准, 2019, 9(6):31-38 Zou W H, Liu L, Sun T, et al. Residues determination and risk assessment of prometryn in aquaculture products[J]. Chinese Fishery Quality and Standards, 2019, 9(6):31-38(in Chinese)
谢剑, 戴习林, 臧维玲, 等. 扑草净对两种虾和两种水草的毒性研究[J]. 湖南农业科学, 2010(23):147-150 Xie J, Dai X L, Zang W L, et al. The toxicity of prometryne on two varieties of shrimp and two varieties of aquatic-weed[J]. Hunan Agricultural Sciences, 2010 (23):147-150(in Chinese)
王田田, 刘丽娟, 孙灵毅. 4种除藻剂对菲律宾蛤仔和刺参的急性毒性实验[J]. 中国渔业质量与标准, 2015, 5(2):35-41 Wang T T, Liu L J, Sun L Y. The acute toxicity of four algaecides to Ruditapes philippinarum and Apostichopus japonicas [J]. Chinese Fishery Quality and Standards, 2015, 5(2):35-41(in Chinese)
田秀慧, 宫向红, 徐英江, 等. 除草剂扑草净在海参中的生物富集与消除效应研究[J]. 现代食品科技, 2013, 29(7):1580-1585 Tian X H, Gong X H, Xu Y J, et al. Accumulation and elimination of prometryn in Apostichopus japonicas [J]. Modern Food Science and Technology, 2013, 29(7):1580-1585(in Chinese)
刘丽娟, 王玮云, 张华威, 等. 菲律宾蛤仔对扑草净的生物富集与消除规律[J]. 食品科学, 2016, 37(21):252-256 Liu L J, Wang W Y, Zhang H W, et al. Accumulation and elimination of prometryn in Ruditapes philippinarum [J]. Food Science, 2016, 37(21):252-256(in Chinese)
张望, 范广宇, 陈秀开. 扑草净在文蛤体内的生物富集与消除规律研究[J]. 水产科学, 2018, 37(3):399-403 Zhang W, Fan G Y, Chen X K. Bioaccumulation and elimination of prometryn in hard clam Meretrix meretrix [J]. Fisheries Science, 2018, 37(3):399-403(in Chinese)
Stará A, Kristan J, Zuskova E, et al. Effect of chronic exposure to prometryne on oxidative stress and antioxidant response in common carp (Cyprinus carpio L.)[J]. Pesticide Biochemistry and Physiology, 2013, 105(1):18-23 Stará A, Kouba A, Velíšek J. Effect of chronic exposure to prometryne on oxidative stress and antioxidant response in red swamp crayfish (Procambarus clarkii)[J]. BioMed Research International, 2014, 2014:680131 黄会, 王共明, 任利华, 等. 扑草净对四角蛤蜊抗氧化酶活性及组织细胞的影响[J]. 中国水产科学, 2021, 28(11):1457-1465 Huang H, Wang G M, Ren L H, et al. Effect of prometryn on the antioxidant enzyme activities and histopathology of Mactra veneriformis[J]. Fisheries Science, 2021, 28(11):1457-1465(in Chinese)
Dong S H, Yang Y Y, Cheng B, et al. Responses of antioxidant defenses in the clam Mactra veneriformis to 2,2',4,4'-tetrabromodiphenyl ether exposure[J]. Comparative Biochemistry and Physiology Toxicology & Pharmacology, 2019, 217:98-105 鲍相渤, 刘卫东, 姜冰, 等. 内参基因在虾夷扇贝定量PCR中表达稳定性的比较[J]. 水产科学, 2011, 30(10):603-608 Bao X B, Liu W D, Jiang B, et al. Expression stability of reference genes for quantitative PCR in Japanese scallop Patinopecten yessoensis [J]. Fisheries Science, 2011, 30(10):603-608(in Chinese)
曹滕飞, 丛明, 李兆艳, 等. 氨态氮对菲律宾蛤仔毒理机制研究的内参基因筛选[J]. 生态毒理学报, 2017, 12(2):182-190 Cao T F, Cong M, Li Z Y, et al. Selection of reference genes for toxicological mechanism research on Ruditapes philippinarum exposed to ammonia nitrogen[J]. Asian Journal of Ecotoxicology, 2017, 12(2):182-190(in Chinese)
常悦. 四角蛤蜊CAT、GPX抗氧化酶基因的克隆[D]. 呼和浩特:内蒙古农业大学, 2013:19-25 栾伟. 中国明对虾重要分子伴侣蛋白基因的克隆及表达研究[D]. 北京:中国科学院研究生院, 2009:70-112 Luan W. Identification and characterization of chaperone protein genes in Chinese shrimp Fenneropenaeus chinensis [D]. Beijing:Graduate School of Chinese Academy of Sciences, 2009:70 -112(in Chinese)
Werck-Reichhart D, Feyereisen R. Cytochromes P450:A success story[J]. Genome Biology, 2000, 1(6):30031-30039 潘鲁青, 宫晓莉, 许超群, 等. 四溴联苯醚对菲律宾蛤仔组织解毒代谢基因表达的影响[J]. 中国水产科学, 2012, 19(2):294-300 Pan L Q, Gong X L, Xu C Q, et al. Effects of tetrabromodiphenyl ether (BDE-47)on the metabolic gene expression in the gills and digestive glands of Ruditapes philippinarum [J]. Journal of Fishery Sciences of China, 2012, 19(2):294-300(in Chinese)
Hayes J D, Strange R C. Glutathione S-transferase polymorphisms and their biological consequences[J]. Pharmacology, 2000, 61(3):154-166 周驰, 李纯厚, 张为民, 等. 翡翠贻贝CYP4基因克隆及其表达水平分析[J]. 热带海洋学报, 2010, 29(4):82-88 Zhou C, Li C H, Zhang W M, et al.CYP4 gene cloning and expression level analysis of Perna viridis [J]. Journal of Tropical Oceanography, 2010, 29(4):82-88(in Chinese)
Behnisch P A, Hoso K, Sakai S, et al. Combinatorial bio/chemical analysis of dioxin and dioxin-like compounds in waste recycling, feed/food, humans/wildlife and the environment[J]. Environment International, 2001, 27(6):495-519 Yu L Q, Lam J C, Guo Y Y, et al. Parental transfer of polybrominated diphenyl ethers (PBDEs) and thyroid endocrine disruption in zebrafish[J]. Environmental Science & Technology, 2011, 45(24):10652-10659 Morel Y, Barouki R. Repression of gene expression by oxidative stress[J]. The Biochemical Journal, 1999, 342(Pt 3):481-496 Takahashi K. Purification and characterization of human plasma glutathione peroxidase:A selenoglycoprotein distinct from the known cellular enzyme[J]. Archives of Biochemistry and Biophysics, 1987, 256(2):677-686 徐雄, 李春梅, 孙静, 等. 我国重点流域地表水中29种农药污染及其生态风险评价[J]. 生态毒理学报, 2016, 11(2):347-354 Xu X, Li C M, Sun J, et al. Residue characteristics and ecological risk assessment of twenty-nine pesticides in surface water of major river-basin in China[J]. Asian Journal of Ecotoxicology, 2016, 11(2):347-354(in Chinese)
张望, 范广宇, 孟祥龙, 等. 海州湾沿岸海水中21种除草剂的分布特征[J]. 江苏农业科学, 2019, 47(23):289-294 Zhang W, Fan G Y, Meng X L, et al. Distribution characteristics of 21 herbicides in seawater along coast of Haizhou Bay[J]. Jiangsu Agricultural Sciences, 2019, 47(23):289-294(in Chinese)
乔丹. 山东沿海贝类中除草剂污染特征及风险评价[D]. 上海:上海海洋大学, 2017:23-37 Qiao D. Characteristics and risk assessment of herbicides residue in coastal shellfish in Shandong Province[D]. Shanghai:Shanghai Ocean University, 2017:23 -37(in Chinese)
郑磊, 杨文龙, 董亮, 等. 扑草净水环境质量基准及风险评估[J]. 中国环境科学, 2021, 41(8):3825-3831 Zheng L, Yang W L, Dong L, et al. Derivation of water quality criteria and ecological risk assessment for prometryn[J]. China Environmental Science, 2021, 41(8):3825-3831(in Chinese)
-
点击查看大图
计量
- 文章访问数: 1016
- HTML全文浏览数: 1016
- PDF下载数: 23
- 施引文献: 0
下载:
